The primary aim of this research is directed towards an understanding of cell wall biosynthesis on the molecular level. In directing our goals, a major emphasis will be placed on the role of the membrane in this process. Knowledge of the membrane-associated enzymes and carriers that are involved in the synthesis of this macromolecular net must be integrated with the control systems that govern the synthesis, assembly, and amplification of this structure. Six specific projects will be investigated: (1) Mechanistic analysis of phospho- MurNAc-pentapeptide translocase: purification and properties of the enzyme. (2) Biosynthesis of spin-labeled peptidoglycan: analysis and interpretation of spectra, isolation, and characterization of products. (3) Biosynthesis of fluorescent-labeled peptidoglycan: development of fluoresent-labeled substrate and synthesis of fluorescent-labeled undecaprenyl-diphosphate-MurNAc-pentapeptide. (4) Biosynthesis of membrane teichoic acid: role of the membrane in the incorporation of D- alanine into teichoic acid. (5) Biosynthesis of peptidoglycan: role of alkyl substituents on the synthesis of nascent peptidoglycan and cross- linked peptidoglycan. (6) Role of the D-alanine-glycine transport system in the biosynthesis of cross-linked peptidoglycan. An investigation of these projects will contribute to our understanding of cell wall biosynthesis and the role of the membrane in this process.